School of Biotechnology, Dublin City University, Dublin, Ireland.
Hamad Bin Khalifa University, Doha, Qatar.
Methods Mol Biol. 2022;2466:3-22. doi: 10.1007/978-1-0716-2176-9_1.
Antibodies are an integral part of many biological assays and biotherapeutics. However, the sources from which antibodies are derived frequently contain other contaminants which may interfere with assays or cause adverse reactions if administered in vivo. Therefore, a means of isolating these antibodies from their source at high levels of purity is critical. Affinity chromatography is currently one of the most widely applied methods for the purification of antibodies. This method relies on specific and reversible, interactions between antibody structures, or recombinant tags fused to these structures, and ligands immobilized on solid support matrices, generally within a column. Herein, common chromatographic methods applied to antibody purification are described. These include the purification of IgG, and its recombinant forms, through protein A, protein G and immobilized metal affinity chromatography.
抗体是许多生物检测和生物疗法的重要组成部分。然而，抗体的来源经常含有其他污染物，如果在体内给药，可能会干扰检测或引起不良反应。因此，需要一种从其来源中以高纯度水平分离这些抗体的方法。亲和层析是目前应用最广泛的抗体纯化方法之一。该方法依赖于抗体结构或与其融合的重组标签与固定在固体支撑基质上的配体之间的特异性和可逆相互作用，通常在柱内进行。本文描述了应用于抗体纯化的常见色谱方法。这些方法包括通过蛋白 A、蛋白 G 和固定化金属亲和层析从 IgG 及其重组形式中进行纯化。
